猪流行性腹泻病毒S1蛋白在干酪乳杆菌中的分泌表达及免疫原性分析  

Recombinant Lactobacillus casei secretory expression of porcine epidemic diarrhea virus spike protein and the immunogenicity analysis

作　　者：葛俊伟[1] 姜艳平[1] 汪淼[1] 乔薪瑗[1] 徐义刚[1] 唐丽杰[1] 李一经[1] 

机构地区：[1]东北农业大学动物医学学院,黑龙江哈尔滨150030

出　　处：《中国预防兽医学报》2009年第4期256-260,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金(30671574)

摘　　要：将猪流行性腹泻病毒(PEDV)纤突蛋白S1基因片段插入干酪乳杆菌分泌型表达载体pPG-2中,构建了重组表达载体pPG-s1,将其电转化干酪乳杆菌L.casei393,获得了表达PEDV S1蛋白的重组乳酸菌杆菌。重组杆菌诱导后,经western blot、间接ELISA实验表明,目的蛋白获得了分泌表达。将该重组干酪乳杆菌经口服接种途径免疫BALB/c小鼠,免疫后于不同时间分别测定了粪便中特异性的sIgA和血清中IgG;用MTT法检测免疫小鼠细胞脾淋巴细胞增殖情况。结果表明该重组干酪乳杆菌表达系统能刺激动物黏膜免疫应答和系统免疫应答,在肠道可产生分泌型IgA抗体,并可检测到高水平血清IgG;不仅能诱导体液免疫反应,还可诱导细胞免疫应答。表明所构建的重组干酪乳杆菌表达系统作为口服疫苗具有潜在的应用价值,为探索新型猪流行性腹泻口服疫苗的研制奠定了基础。To evaluate the immune responses of recombinant L.casei 393 expressing porcine epidemic diarrhea virus （PEDV） S1 protein as a oral vaccine, the spike gene fragment of PEDV was subcloned into the expression vector pPG-2, and transformed into L.casei 393 by electroporation to generate recombinant becteria of pPG-2-sl/L.casei 393. The recombinant bacteria was adminstered to BALB/c mouse via oral administration. The results showed that the mice immunized with recombinant bacteria produced high level of anti-PEDV IgG in the serum and high level of sIgA detected in the feces. Significant differences in the spleen lymphocyte proliferation index （LPI） were observed among the immunization groups of mice and control groups. The serum antibodies had neutralizing effect against PEDV infection. In conclusion, the oral immunizations with recombinant L.casei 393 could induce specific mucosal lgA response as well as serum IgG responses, and evoked both mucosal immune and system immune responses against PEDV.

关 键 词：猪流行性腹泻病毒 S蛋白 干酪乳杆菌 免疫反应 

分 类 号：S852.651[农业科学—基础兽医学]